利用组成型启动子在代谢工程大肠杆菌菌株中高效生产谷胱甘肽。
Efficient glutathione production in metabolically engineered Escherichia coli strains using constitutive promoters.
机构信息
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, 200237, China.
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, 200237, China; Shanghai Collaborative Innovation Center for Biomanufacturing Technology, 130 Meilong Road, Shanghai, 200237, China.
出版信息
J Biotechnol. 2019 Jan 10;289:39-45. doi: 10.1016/j.jbiotec.2018.11.001. Epub 2018 Nov 3.
Glutathione (GSH) is an important bioactive tripeptide and widely used in food, medicine and other industries. Recently, the bifunctional glutathione synthetase, GshF, has been applied for efficient GSH production under inducible promoter. In this study, the constitutive expression of GshF from Streptococcus sanguinis (GshFss) was investigated under four different constitutive promoters. Based on previous study, five genes in Escherichia coli JM109 were deleted to eliminate the degradation of precursors and GSH. The effects of gene knockout on the constitutive expression of GshFss and GSH production were evaluated by whole cell catalysis. Finally, the engineered strain JM03P produced 24 mM glutathione with addition of 30 mM precursors in 5-L bioreactor fed-batch fermentation. The yield of GSH based on cysteine in JM03Pwas reached 80% without any inducer, which was improved by 17.3% than that in the control strain.
谷胱甘肽（GSH）是一种重要的生物活性三肽，广泛应用于食品、医药等行业。最近，双功能谷胱甘肽合酶 GshF 在诱导型启动子下被用于高效生产 GSH。在这项研究中，研究人员考察了来自酿脓链球菌（GshFss）的组成型表达 GshF 在四种不同组成型启动子下的情况。基于先前的研究，敲除了大肠杆菌 JM109 中的五个基因，以消除前体和 GSH 的降解。通过全细胞催化评估基因敲除对 GshFss 的组成型表达和 GSH 生产的影响。最后，工程菌株 JM03P 在 5-L 生物反应器分批补料发酵中添加 30mM 前体可生产 24mM 谷胱甘肽。与对照菌株相比，JM03P 中基于半胱氨酸的 GSH 得率提高了 17.3%，无需任何诱导剂即可达到 80%。
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